C CI)r Collection anfc 
pcraertoatton of fepect 
mens for laioratorp 
Cjramtnatton 



Herbert C. Emerson, M.D. 



The Collection and Preservation 
Specimens for Laboratory 
Examination 



The 

Collection and Preserva 
tion of Specimens for 
Laboratory Ex 
amination 



By 

Herbert C. Emerson, A.B., M.D. 

Laboratory, 177 State Street 
Springfield, Massachusetts 
Telephone 543 



Copyright 1904 
By Herbert C. Emerson 



Tv?t> tones h*e**»v«d 
[ OCi 17 1904 
I . &>&yrteht £mrv 

\ CL&SS <^_XXo. No, 1 

; OOPY b 



INDEX 

Blood 17 

Examination for Leucocytosis . . 17 

Examination for Malaria .... 20 

Examination for Typhoid Fever . . 21 

Examination for other abnormal conditions 17 
Chemical Analyses — Foods, Papers, Cloths, 

Gas, etc . . 29 

curettings 25 

Diphtheria 14 

Gonorrhoea 15 

Ice . 32 

Intestinal Contents . . . . .24 

Malaria 20 

Memoranda, Blank Forms for 33 

Milk — Cow's 29 

Woman's 28 

Secretions and Exudations ... 27 

Specimens, Suggestions for Sending . . 8 

Sputum 12 

Stomach Contents 23 

Test Breakfast 23 

Tumors 25 

Typhoid Fever, Widal Test for . . 21 

Urine .10 

Water . -30 



5 




iHE object of this booklet is 
to serve the interests of phy- 
sicians who are desirous of 
having certain laboratory 
tests made, and who, for lack 
of time or equipment, have not the oppor- 
tunity to make such examinations them- 
selves. 

No laboratory worker desires the results 
of his examinations to be considered inde- 
pendent of the clinical conditions, but 
rather in conjunction with, and supple- 
mentary to, the history, and physical signs 
and symptoms of the patient. A skeleton 
history of the case is, therefore, essential 
to the laboratory investigator to properly 
interpret the results of his analyses in giv- 
ing an opinion on the case. 

It is the hope of the writer that the 
following pages may facilitate the inter- 
dependence of the clinician and the lab- 
oratory worker, by suggesting some points 
of practical utility in the preparation of 
specimens intended for examination in the 
laboratory. 

7 



General Considerations 



For the satisfactory examination of any 
material it is essential that the specimen 
should be received in proper condition, so 
that the analysis may truly determine what 
is desired by the sender. To this end the 
laboratory is prepared to furnish suitable 
outfits and receptacles for all sorts of spec- 
imens as hereinafter specified. Sugges- 
tions for the proper preparation of the 
various kinds of material accompany each 
outfit, and as these outfits are furnished 
without expense the laboratory is not only 
willing, but desirous, to forward them at 
any time on request. 

Whatever assists the laboratory worker 
in the examination of the specimen con- 
tributes to the value of the report to the 
sender, and in order to be satisfactorily 
handled at the laboratory, all material for 
examination should be accompanied by a 
statement of the following facts: 

i. Name and address of person to 
whom report is to be sent. 

8 



2. Name, age and sex of patient, (for 
laboratory record only). 

3. Source and nature of the specimen. 

4. Object of examination. 

Blank forms for the above information 
are sent with all outfits. 

Any clinical facts which may contribute 
to a better understanding of the case will 
be of interest and service to the examiner. 

Written reports are sent as soon as the 
examination is completed. 

Reports by telephone or telegraph are 
sent when so requested. 

Packages should be sent by express, 
securely wrapped, plainly marked, and a 
"Rush" tag attached. 



9 



Urine 



In order that proper inferences may be 
drawn from an examination of urine, it is 
essential that the specimen be collected in 
the following manner: 

All the urine passed in 24 hours, includ- 
ing that voided at stool, should be saved 
in a clean, covered receptacle kept in a 
cool place. It is best to select some hour 
for beginning the collection, preferably in 
the morning, say at 7 o'clock, and save all 
urine passed up to that time on the follow- 
ing morning. The total quantity should 
be carefully measured, and no less than 
four ounces of the mixture sent for the 
analysis. It is especially important that 
a record of the total twenty-four hour 
amount be sent with the sample. 

The report of an examination of urine 
embraces the qualitative and quantitative 
chemical tests, and the microscopical exam- 
ination. When special tests are to be made 
like examination of the sediment for tuber- 
cle bacilli, gonococci, etc., a request for the 
10 



same should accompany the specimen. 
Very often the physician is in possession 
of facts in the history of the case that 
have important bearing on the character 
of the inferences that may be drawn from 
the results of the laboratory examination. 
Such facts should be submitted with the 
sample of urine. 

Urine collected in the warm months 
rapidly undergoes decomposition on stand- 
ing, and it is therefore advisable to add 
to the sample a few drops of formalin, or 
salicylic acid solution. 

Four-ounce bottles in suitable mailing 
cases are furnished by the laboratory. 



it 



Sputum 

Examination of material raised from the 
air tubes and lungs furnishes valuable in- 
formation as to the cause of pulmonary 
and bronchial affections. 

Evidences of such morbid conditions 
are varied in character, and include many 
pathological findings in addition to the 
presence of bacteria such as tubercle ba- 
cilli. Tubercle bacilli may often be dem- 
onstrated in the blood from pulmonary 
hemorrhage. 

Sputum should be collected in wide- 
mouthed, clean bottles. Only such spu- 
tum as is coughed up from the air tubes 
and lungs should be deposited in the bottle. 
Material coming from the mouth contain- 
ing saliva and mucus and that coughed 
up directly after eating are not suitable 
for examination and should be rejected. 
Sputum which is coughed up before the 
morning meal is often the most satisfac- 
tory for examination. The amount re- 
quired is not large, but where the total 



12 



daily quantity expectorated is small, one 
or even two days' expectoration should be 
reserved. 

As there is danger to the examiner in 
handling tubercular sputum, care should 
be taken that the material be prevented 
from leaking from the receptacle. Cork 
stoppered bottles only should be used. 
Screw -capped bottles are dangerous to 
handle and should never be used for spu- 
tum. 

Wide-mouthed bottles without necks in 
cases suitable for transportation are fur- 
nished by the laboratory. 

If for any reason a specimen is unsat- 
isfactory a second will be requested. 



13 



Diphtheria 

This laboratory is prepared to make the 
necessary examinations for the diagnosis 
of diphtheria in suspected cases, and for 
the examination of release cultures in cases 
about to be discharged from isolation, for 
individuals or for local boards of health. 

Suitable outfits containing sterile swabs 
and tubes of culture media are furnished 
by the laboratory. Reports are made as 
soon as possible by telephone, telegraph 
or letter, as the sender desires. Outfits 
when returned should be sent by express, 
as arrangements are made for immediate 
delivery of such packages. 



14 



Gonorrhoea 



Examination of a discharge for gonococci 
is made by the study of a thin film of the 
secretion, after appropriate staining. 

Pus, suspected of containing gonococci, 
which has been collected on cotton, or on 
a swab, does not furnish satisfactory ma- 
terial for examination, as the secretion 
becomes dried before reaching the labora- 
tory. 

In order to secure specimens suitable 
for examination the following procedure 
is necessary: 

In all cases great care should be taken 
that none of the surrounding tissue be 
touched while obtaining the specimen. 

Secure a drop or two of the discharge 
on a swab and rub well over one side of a 
glass slide. Two or more slides should be 
prepared. 

In cases in which the discharge is to 
be taken from the os uteri or the female 
urethra, the swab wire may be held in a 
pair of artery or other forceps, avoiding, 
J 5 



as far as possible, contact with the vaginal 
walls. 

The slides will dry quickly in the air 
and should then be placed in a box and 
sent to the laboratory. 

The outfit furnished by the laboratory 
for the preservation of a gonorrhoeal dis- 
charge consists of a wooden box, contain- 
ing a sterile swab for securing the material, 
and slides upon which the smear should 
be made. The package should be sent 
by express. 



16 



Blood 



An accurate enumeration of the number 
of red and white blood corpuscles can only 
be made by counting the cells themselves, 
and this necessitates the taking of the sam- 
ple of blood by means of special apparatus 
which demands the attendance of the ex- 
aminer. Personal visits are always gladly 
made, however, for this purpose. 

Aside from the actual counting of the 
blood corpuscles, much valuable informa- 
tion as to pathological conditions of the 
blood can be obtained from a study of a 
stained thin film of blood smeared upon 
glass slides. 

Briefly, the following facts may be de- 
termined: 

Presence of malarial or other organisms. 

Approximate estimation of the amount 
of haemoglobin and the ratio of red and 
white cells. 

Presence of abnormal forms of red and 
white cells. 

Differential count of white cells. 
J 7 



Directions for Preparing Specimens 
of Blood 

1. Cleanse the skin of the lobe of the 
ear with soap and water. Dry. 

2. Prick the lobe deeply to insure free 
escape of blood. 

3. Avoid the first few drops; touch the 
escaping blood with the surface of a slide 
about one-half inch from the end, to secure 
a drop of blood. 

4. Place the slide on some firm object, 
as table or book. 

5. Take a second slide by the edges 
between thumb and fingers, allowing the 
other end to rest just ahead of the drop 
of blood on the first slide used. 

6. Holding the slide at about an angle 
of 45 degrees, draw it slowly but with firm 
and even pressure down the length of the 
slide, thus distributing the blood in a thin 
layer over the surface. 

7. Repeat this procedure, using the 
slide first smeared with blood as the 
"spreader." 

18 



8. Allow the slides to dry in the air, — 
when they are ready to be sent to the 
laboratory. 

In handling glass slides preparatory to 
making blood smears, too great care can- 
not be taken to avoid touching the surface 
of the slides by the fingers. It is well to 
wash the slides in strong alcohol just before 
using, wiping dry with old linen. 

Outfits and directions for preparing the 
smear are furnished by the laboratory, and 
when returned, should be accompanied 
with a statement as to the object of the 
examination. 



19 



Malaria 

To determine the presence of malarial or- 
ganisms, it is necessary to study a fresh 
smear of blood, or a stained smear of dried 
blood. The latter method alone is appli- 
cable to specimens coming from a distance, 
and directions for preparing the smears 
are to be found on page 18 under the cap- 
tion "Blood." 



2C 



The Widal Test 

Serum Diagnosis of Typhoid Fever 

"This method of diagnosis is based on 
the fact that living and actively motile 
typhoid bacilli if placed in a diluted serum 
of a patient suffering from typhoid fever, 
within a short time lose their motility and 
become aggregated into clumps." 

This test is carried out by means of a 
fresh bouillon culture of typhoid bacilli in 
conjunction with a watery solution of dried 
blood. It should be noted: 

A, That a positive reaction is not con- 
clusive evidence of the existence of typhoid 
fever if the patient has had the disease 
before, but for all practical purposes a 
positive result may be taken to indicate 
the existence of this disease. 

B. That a single negative result, un- 
less obtained after the seventh or eighth 
day, has little value, and repeated tests 
should be made in negative cases. 

The method used in this laboratory for 

performing the Widal test consists in using 
21 



the dried blood, which is best collected in 
drops (not smears) on glass or non-ab- 
sorbent paper like a visiting card. The 
blood is allowed to dry in the air, and is 
then ready to be sent to the laboratory. 

Secondary examinations are often nec- 
essary to establish a satisfactory diagnosis 
and are always gladly made. 

As this method of diagnosis involves 
the use of a diluted blood serum, such 
accuracy as is possible in collecting the 
drop should be observed. To this end the 
outfit furnished by the laboratory consists 
of a glass slide, a needle for puncture and 
a wire loop to be used for collecting the 
drop. This outfit may be sent by mail. 



L.of C. 



Stomach Contents 



The differentiation between organic and 
functional diseases of the stomach is great- 
ly helped by a chemical and microscopic 
examination of the gastric contents. Pos- 
itive evidences of new growths may be 
found and variations in the degree and 
kinds of acidity, and indications of imper- 
fect digestion. 

The most satisfactory examination of 
the contents of the stomach is made when 
the contents have been expressed after a 
test meal. The test breakfast of Ewald 
and Boas is recommended, consisting of 
one or two white rolls, one large cup of 
tea (without milk or sugar) or water. 

This should be taken while the patient 
is fasting, preferably in the morning before 
anything is taken into the stomach, and the 
stomach contents expressed or secured by 
the use of the stomach pump one hour 
after the meal. The contents should be 
collected in a clean bottle and sent to the 
laboratory as soon as possible. 

23 



Intestinal Contents 



Examination of faecal discharges for the 
presence of blood, shreds of new growths, 
tubercle bacilli, etc., are most satisfactorily 
made when the material can be freshly 
examined. The stool should be collected 
in a clean wide-mouthed bottle, and should 
be sent as soon as possible to the labora- 
tory. 

A history of the case with the object 
desired in the examination should be 
stated. 



24 



New Growths — 

Curettings 

Place the tissue in either weak alcohol, 
or preferably weak formalin solution. 

Special care is to be taken that the 
tissue does not become dry. 

The material for examination should 
be sent to the laboratory as soon as pos- 
sible after removal. 

When the tissue is excised for the pur- 
pose of diagnosis, as large a piece as pos- 
sible should be removed, and the point of 
excision carefully chosen so as to embrace 
not only the superficial but the deeper por- 
tions of the suspected new growth. In 
case a uterine curetting is to be sent for 
examination, special pains should be taken 
to secure scrapings sufficiently deep to dem- 
onstrate invasion of the body of the uterus, 
and all the tissue removed should be pre- 
served. 

In sending specimens of this character 
for examination it is essential that a brief 



25 



history of the case be also submitted, as 
well as the source of the specimen. Tis- 
sue of a certain kind may be normal when 
found in one place in the body and path- 
ological when appearing elsewhere. 



26 



Secretions, Exudations, 
etc. 

Material of this character is best spread 
on glass slides, as soon as taken from the 
patient. When collected on swabs it is 
apt to become dry and useless for tests 
unless sent directly to the laboratory in 
sealed test-tubes. 

When cultures are to be made from 
such pathological material, sterile swabs 
must always be used and care taken that 
no disinfectant is used just previous to 
taking the specimen. In securing a swab 
of a discharge from the ear, for example, 
sterile water should be first used to cleanse 
the cavity. 

Fluids from the chest and abdominal 
cavity should be collected in sterile bottles 
which should be tightly stoppered with 
boiled corks. 



27 



1 



Milk 

Examinations of mother's milk for the 
percentage of fat, sugar and protein can 
be satisfactorily made with a sample of 
middle milk from both breasts. Two 
ounces should be sent as the sample. 



28 



Chemical Analyses 

Aside from clinical laboratory work, qual- 
itative chemical analyses are made at this 
laboratory, determining the presence of the 
various elements in such mixtures as alloys, 
coal, paper, air, gas, flue gas, etc. Quan- 
titative determinations of such substances 
will also be made. 

In addition to inorganic analyses, the 
laboratory is prepared to make analytical 
determinations of organic compounds. 

By way of illustration, the following 
analyses may be mentioned: analysis of 
various liquors for adulterants and deter- 
minations of alcohol, examination of water, 
cow's milk, cream, butter and other foods, 
wall paper and other material for arsenic. 



29 



Water 



Satisfactory chemical analyses of drink- 
ing water are possible only when the sample 
to be examined is carefully collected in 
perfectly clean glass bottles holding at least 
one gallon, with glass stoppers or new 
corks. Such bottles in proper cases for 
shipping are furnished. 

Directions for Taking the Sample 
of Water 

1. From a pump or tap. 

Allow 7 w^ater to run freely for two min- 
utes before sample is taken. 

Rinse bottle three times and fill. 

Rinse stopper and put in place. 

Secure stopper by tying a clean piece 
of cotton cloth over it, bringing the ends 
of the string up over top of stopper where 
they should be sealed with wax. 

2. From stream, or open well. 

If collected from stream or well avoid 
getting sediment into sample. 

30 



Avoid the scum on the surface. 

Rinse bottle and stopper, and fill bottle 
below surface. 

If the water is not deep enough to do 
this, the water may be dipped up by an 
absolutely clean vessel, and poured into 
bottle after it has been rinsed. 

The sample of water should be collected 
immediately before shipping, so that as 
little time as possible shall intervene be- 
tween the collection of the sample and its 
examination. 



Bacteriological examinations of water 
to determine the presence of pollution are 
often more valuable than the chemical tests, 
while the examination of water by both 
methods is frequently desirable. For the 
bacteriological examination the sample 
must be collected in water bottles, ster- 
ilized for the purpose, to be obtained at 
the laboratory. In many cases a personal 
inspection of the region about the source 

3i 



of the water to be tested is a most essen- 
tial aid in giving an opinion as to the 
character of the water. 

Microscopical examination of water for 
the kind and number of vegetable and 
animal organisms are made at this lab- 
oratory; also examinations of ice for evi- 
dences of contamination and tests for the 
bacterial efficiency of filters. 



32 



Memorandum to be sent with Specimen 



Name Age 

Male 
Female 



Source of Specimen 



Object of Examination 



History 



Report to 



Memorandum to be sent with Specimen 



Name Age 

Male 
Female 

« 

Source of Specimen 



Object of Examination 



History 



Report to 



Memorandum to be sent with Specimen 



Name Age 

Male 
Female 

Source of Specimen 



Object of Examination 



History 



Report to 



